转录因子CsAGL6和 CsERF110在柑橘色泽等品质协同变化中调控作用的研究

It is known that the formation of fruit qualities was not isolated, but come up as the comprehensive performance of a number of individual qualities, which influence and restrict each other as a whole. This may be derived from the regulation of some common regulatory genes shared in different nodes of different metabolic pathways. Transcription factors are important regulatory proteins, playing regulator roles in a wide range of plants growth and development processes. From early studies, we found potential transcription factors such as CsAGL6 and CsERF110 in citrus as Arabidopsis homologous, which affected carotenoids metabolisms, and simultaneously imposes significant effects on the formation of other qualities, with its regulation mechanism unclear. On this basis, the project intends to overexpress genes CsAGL6 and CsERF110 in citrus calli and tomato, respectively, then the obtained transgenic materials will be subjected to ChIP-seq, RNA-seq and yeast one hybrid analyses to detect and verify target genes regulated by the two transcription factors; at the same time, metabolomics method will be applied to reveal the synergistic regulation of transcription factors on main qualities related metabolites such as pigments and aroma; Furthermore, combined evidences from metabolomics and molecular biology will shade light on the mechanisms of how the transcription factors synergistically regulate the fruit qualities of pigmentation and others. The results will benefit the research on the synergy regulation mechanism of several qualities, as well as the field practices to improve fruit integrated qualities by providing theoretical basis.

果实品质不是孤立的，而是多个单项品质的综合表现，它们在形成过程中相互影响和制约，可能与多个品质相关代谢途径的节点受共同的基因调控有关。转录因子是一类对生长发育有重要调控作用的蛋白质。我们前期挖掘到与拟南芥同源的柑橘转录因子CsAGL6和CsERF110，并初步明确它们除与类胡萝卜素代谢有关外，还影响其他品质的形成，但其调控机制尚不清楚。基于此，本项目拟将CsAGL6和CsERF110基因分别在富含类胡萝卜素的柑橘愈伤组织和番茄中超量表达，并将获得的转化材料，采用ChIP-seq、RNA-seq和酵母单杂交等技术寻找并验证其所调控的靶基因；同时，利用代谢组学方法分析该转录因子对色泽、香味等主要品质相关物质代谢的协同调控；进一步，结合代谢组学和分子生物学证据来解析柑橘果实色泽与多项品质的关联性形成机理。本研究结果，对于探讨多项品质间的协同变化机制，进而寻找提升果实综合品质的方法提供了理论依据。

果实品质是多个单项品质的综合表现，它们相互影响和制约，可能与多个品质相关代谢途径的节点受共同的基因调控有关。转录因子是一类对生长发育有重要调控作用的蛋白质。我们针对前期采用组学技术挖掘到的4个柑橘转录因子CsAGL6、CsPIF8、CsERF110和CgMYB58，在明晰柑橘中类胡萝卜素、木质素及叶绿素的代谢规律和种质特异性的基础上，采集有极端表型的不同种质，分别采用转录组测序、染色质免疫共沉淀—高通量测序、双荧光素酶、WGCNA分析、凝胶阻滞、CoIP-质谱分析、基因转化、柑橘汁胞离体培养和外源激素处理等技术，解析了CgMYB58对华农红柚汁胞中木质素及类胡萝卜素的共同调控机理；明确了CsAGL6和CsPIF8对甜橙等柑橘果实中品质相关多个代谢途径的协同调控；揭示了响应外源ABA的CsERF110对柑橘不同组织中类胡萝卜素合成的调控。研究结果在丰富了柑橘汁胞粒化及类胡萝卜素调控机理基础理论的同时，加强了研究者对多代谢途径协同调控的认知，也为产业实践上抑制柑橘汁胞木质化及提高柑橘综合品质提供了重要的理论依据。